(9/7/05) The following is a list of programs on the Web that I find
useful for analyzing DNA and designing cloning strategies.
Primer3:
I've found this to be a simple way to identify primer pairs for PCR
amplification. Its particularly useful for identifying a pair of
primers for cloning PCR fragments because you can set the sequence of
one primer or constrain the region in which one or both primers are
sought.
IDT SciTools:
We buy most of our oligos from IDT. This link provides a primer
selection program that I've not used - I use Primer3. I use OligoAnalyzer
3.0 at the IDT site to make certain primer combinations are not
likely to form homo- or hetero-dimers.
NEBcutter v2.0:
This program identifies restriction sites and open reading
frames. I've found it very useful for designing cloning
strategies that require the fusion of two open reading frames.
The program allows you to do a BLAST analysis on any of the open
reading frames. This is useful for identifying the proteins in
the clone and for locating junctions between fusion proteins.
Local Alignment Tool:
This program aligns matching sequences. I
like it because it retains the numbering of bases used in a DNA strider
file so I can easily go back and forth. Its useful when trying to
align real sequence results with a sequence in strider or for locating
where an oligo makes a complete or partial match to another sequence.
FlyBase: This is your
entry to a wealth of information for working with Drosophila.